JPS6281322A - Aldose reductase inhibitor - Google Patents

Abstract

PURPOSE:To provide the titled inhibitor composed of GYUSHA-JINKIGAN (a kind of compounded Chinese herb drug). CONSTITUTION:A mixture composed of 5-8pts.(wt.) of JIOU (root of Rehmania glutinosa), 2-3pts. of GOSHITSU (root of Achyranthes fauriei), 2-4pts. of YAMAGUMI, 3-4pts. of SANYAKU (rhizome of Dioscorea japonica), 2-3pts. of SHAZENSHI (seed of Plantago major), 3pts. of TAKUSHA (tuber of Alisma orientale), 3-4pts. of BUKURYO (Pileus of Poria cocos), 3pts. of BOTANPI (root skin of Paeonia moutan), 1-2pts. of KEIHI (bark of Cinnamomum cassia) and 0.5-1pt. of BUSHI (tuber of Aconitum carmichaeli) is extracted with 10 times weight of hot water and the extracted liquid is filtered and dried to obtain dried extract powder of GYUSHA-JINKIGAN useful as an aldose reductase inhibitor. It can be administered in the form of powder, granule, tablet, capsule, etc. If necessary, the extract may be purified by dialysis or various chromatographic means. Dose: 1-10g daily in about 3 divided doses. It has extremely low toxicity and high safety.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an aldose reductase inhibitor comprising Goshajinkigan.

In recent years, the accumulation of intracellular sorbitol via the polyol pathway, which is a glucose metabolic pathway, has attracted attention as a cause of various complications of diabetes such as cataracts, retinopathy, neuropathy, and nephropathy. The polyol pathway is a metabolic pathway in which aldoses such as glucose and galactose are converted to ketoses such as fructose via polyols such as sorbitol and galactitol, and immunohistochemical techniques have revealed that they are widely present in all organs throughout the body. It has become.

The first step in this pathway, the enzyme that catalyzes the conversion between aldose and polyol, is called aldose reductase.
This enzyme is considered to be the rate-limiting enzyme in the polyol pathway. It has been reported that inhibiting this aldose reductase and reducing the production and accumulation of sorbitol is effective in treating complications in diabetic patients.

Therefore, the present inventors conducted research on the aldose reductase inhibitory effects of various Chinese herbal prescriptions, and as a result, found that the results of research on the aldose reductase inhibitory effects of various Chinese herbal formulas were as follows. It has been discovered that a Chinese herbal medicine formula consisting of , Keirin, Mutanpi, Cinnamon, and Fuzi, that is, Goshajinkigan, has an aldose reductase inhibitory effect. The present invention is based on this knowledge and provides an aldose reductase inhibitor consisting of Goshajinkigan, which is effective in treating complications in diabetic patients. Gosha Jinkigan is described in the classic Chinese medicine prescription (Saiseikata), including its constituent herbal medicines, preparation, extraction method, etc., and is used for various diseases such as lower limb disease, back pain, urinary difficulty, and frequent urination. ing. However, it was not previously known that it had an aldose reductase inhibitory effect.

Gyushurajinkigan is made according to the classics, with 5g of daikon, 3g of gyukichi, 3g of wild plants, 3g of wild herbs, 3g of gurumaenji, 3g of swamp, and 3g of ketsurei.
, 3 g of peony bark, 1 g of cinnamon bark, and 1 g of Fuzi can be decocted with 300 liters of water to make 180-g, which can be taken as an aldose reductase inhibitor. Chinese herbal medicine extracts can be used as aldose reductase inhibitors. For example, 5 to 8 ton of rhizomes, 2 to 3 ton of oxtail, 2 to 3 ton of wild plants. 1 part x parts, 3 to 4 parts by weight of wild herbs, 2 to 3 parts by weight of chamianko, 3 parts by weight of sagebrush, 3 to 4 parts by weight of ketrei, 3 parts by weight of peony bark, 1 to 2 parts by weight of cinnamon, 0 parts by weight .5 to 1 part by weight lo(g
After hot extraction with im water, the resulting extract was filtered and dried to obtain a powder of the dried extract of Gyu'kochigan, which is an aldose reductase inhibitor. Add agents, adjuvants, etc. and follow the usual method of manufacturing formulations.
It can also be used in preparations such as powders, granules, tablets, and capsules. If desired, this extract may be further purified and used by conventional methods such as dialysis and various chromatography.

A specific example of the production of the aldose reductase inhibitor of the present invention is as follows.

Specific examples: 5g of rhizome, 3g of cow knees, 3g of mountain radish, 3g of mountain medicine, 3g of Kurumaenji
Add 10 times M1, or 280-, water to a mixture of herbal medicines of g, 3 g of Sawatan, 3 g of Hagirei, 3 g of Mudanpi, 1 g of Cinnamon, and 1 g of Fushi, heat and extract at about 100 ° C. for 1 hour, and the obtained extract was extracted with I. It was retired and spray dried to obtain 3.2 g of dry extract powder.

Next, the fact that the aldose reductase inhibitor of the present invention has an aldose reductase inhibitory effect will be explained with reference to experimental examples.

Experimental Example 1 <Measurement of aldose reductase activity> A 6-year-old male Wistar rat was sacrificed under ether anesthesia, and the crystalline lens was immediately removed and stored at -20°C.

The crystalline lens was homogenized in 135mM sodium-potassium-phosphate buffer (pI+7,0) containing 0.5mM phenylmethylsulfonyl fluoride.
Centrifugation was performed at 00 rpm for 30 minutes. The supernatant was used as a sample for aldose reductase activity measurement.

Furthermore, all of the above operations were performed at 4°C, and the specimens were stored at 0°C.

Aldose reductase activity was measured using Dufuran (Dufuran).
frane) et al. [Biochemical Me.
dicine.

32.99-105 (1984)].

i.e. 100mM lithium sulfate, 0.03+++M
N A D P 11 (reduced nico Linami
de adenine dinucleotide ph
osphate), and 0.1mM DL- as a substrate.
Sample 10 of the above sample was added to 800ml of 135mM sodium-potassium-phosphate buffer (p117.0) prepared to contain glyceraldehyde or 20mM glucose.
0 [and the drug obtained in the above specific example in distilled water at 1 mg/m
A drug solution solution dissolved to give ρ was added to each solution, and the mixture was reacted at 30° C. for 30 minutes. Next, 0.5
The reaction was stopped by adding 0.3-N hydrochloric acid, and by adding 6N sodium hydroxide I- containing 10 mM imidazole, the NADP (oxidized form ni'' cotinamide adenine din
ucleotidepHosphate) was converted into a fluorescent substance, and its fluorescence intensity was measured 60 minutes later. The fluorescence intensity was determined using a spectrophotometer RF-510 (manufactured by Shimadzu Corporation) at room temperature with an excitation wavelength of 360 nm and a fluorescence wavelength of 46 nm.
It was measured under the condition of 0 nm. Further, the fluorescence intensity measured by reacting in the same manner as above except that distilled water was added instead of adding the drug solution was used as a control value.

Aldose reductase is an enzyme that converts DL-glyceraldehyde or glucose into polyol using NADP 11 as a coenzyme, and with this reaction, NADP II is converted to NADP. Therefore, if NADP is low, aldose reductase is inhibited.

As a result, the degree of inhibition of the aldose reductase activity of rat lenses by the drug obtained in the specific example was 72.6% when the substrate was DL-glyceraldehyde, and 38.7% when the substrate was glucose. there were.

From the above results, it was confirmed that the aldose reductase inhibitor of the present invention inhibits the activity of aldose reductase, and is expected to be effective in preventing or treating complications of diabetes.

Next, we conducted an acute toxicity test for oral administration of the drug obtained in the specific example.
When conducted using dY mice and Wistar rats, the drug obtained in the specific example weighed 15 g/
There were no deaths after oral administration of 100 kg (dose limit).

As described above, the drug of the present invention has extremely low toxicity and high safety.

Considering the experimental data and the results of acute toxicity tests in the present invention, the effective dosage m of the drug of the present invention varies depending on the patient's age, weight, and degree of disease, but is usually 10 mg as a dry extract powder for adults. It is considered appropriate to take 1-10g of the drug in three divided doses a day depending on the symptoms.

Next, the present invention will be specifically explained using an example, but the present invention is not limited thereto.

Example 1 200g of the drug produced according to Example 1 above was mixed with 95g of lactose.
g and 5 g of magnesium stearate, and this mixture was compression molded, then ground, sized, and sieved to obtain good granules of 20 to 50 mesh particles.

This granule is suitable for U゛11.5 to 1 ff per day depending on the symptoms.
Take 5 g (corresponding to 1-10 g of dry extract powder of the aldose reductase 1+n inhibitor of the present invention) in three doses.

Example 2 200 g of the drug produced in Example 1 above was mixed with 45 g of microcrystalline cellulose and 5 g of magnesium stearate, and this mixture was compressed using a single-shot double-key press to produce tablets with a diameter of 9 mm and a weight of 1250 mg. . One tablet contains 200 mg of the drug of the present invention.

This tablet is a daily dose of 5 to 50 tablets 3 to 6 times depending on the symptoms.
Take in divided doses.

Example 3 250 g of the drug produced according to Example 1 above was filled into hard capsules. This capsule is administered 4 times a day depending on the symptoms.
Take ~40 capsules in 3 to 6 divided doses a day.

Claims (1)

[Claims] An aldose reductase inhibitor consisting of Goshajinkigan.